A fungal extracellular pectinase composition and its application

Abstract

The present invention provides a fungal extracellular pectinase composition and its application, belonging to the technical field of pectinase. The fungal extracellular pectinase composition obtained by the present invention is composed of 50-100 parts of pectinase composite particles, 0-10 parts of cellulase, 5-16 parts of acid amylase, and 0.5-1.5 parts of surfactant; the pectinase composite particles are prepared by loading liposomes encapsulating fungal extracellular pectinase in chitosan gel. The multiple enzymes in the fungal extracellular pectinase composition obtained by the present invention have a synergistic effect, a fast decomposition speed of pectin, and the prepared pectinase composite particles have high activity, high reuse rate, high environmental tolerance, and high activity at higher temperatures and low pH; the preparation method is simple, can be naturally degraded, non-toxic, and suitable for industrial production.

Description

Technical Field

[0001] The invention belongs to the technical field of pectinase, and particularly relates to a fungal extracellular pectinase composition and application thereof. Background Art

[0002] Pectin, also known as polygalacturonic acid (PG), has a basic polygalacturonic acid skeleton formed by α-1,4 glycosidic bonds. The main chain is polymerized by galacturonic acid (PGA), and a small amount of galactose, arabinose, xylose and rhamnose polysaccharides are connected to the main chain on the side chain. According to the different modification types of the pectin main chain, pectin can be divided into four categories. Among them, protopectin is insoluble in water, and it can produce pectin and pectic acid after hydrolysis; pectic acid is water-soluble, and it is a galacturonic acid polymer that is almost not methylated; when the degree of methylation of polygalacturonic acid reaches 0-75%, it is called pectin ester acid, and when the degree of methylation reaches more than 75%, it is called pectin. According to the different mechanisms of action of pectinase, pectinase can be divided into protopectinase, pectinesterase (PE) and depolymerizing enzymes. Depolymerase can be further divided into hydrolase (Polygalacturonase, PG) and pectin lyase (Pectin lyase, PL). Hydrolase is polygalacturonase, which breaks the α-1,4 glycosidic bonds of the pectin polymer main chain by hydrolysis, while lyase breaks the pectin polymer main chain by trans-elimination and generates unsaturated bonds at the non-reducing end.

[0003] Pectinase can be used in the pre-treatment process of extracting bioactive substances from plants, and is widely used in juice clarification, hemp degumming, textile industry, papermaking, feed, environmental protection and sewage treatment industries. In addition, the application of pectinase in fruit peeling also has good prospects. It can reduce the energy consumption of pulp during peeling, reduce wastewater discharge, and improve product safety.

[0004] Although pectinase has been widely used, there are problems such as difficulty in separation and recovery, low reuse rate, and poor environmental tolerance when using pectinase. Therefore, the enzyme is usually fixed on the carrier particles by enzyme fixation to make it easy to separate and increase the number of enzyme reuses due to the protective effect of the carrier. However, there are still problems such as poor fixation ability, low enzyme activity after fixation, and long enzymolysis time. Summary of the invention

[0005] In view of this, the object of the present invention is to provide a fungal extracellular pectinase composition and its application. The fungal extracellular pectinase composition prepared by the present invention comprises pectinase and auxiliary enzymes, which have a synergistic enzymatic effect and improve the enzymatic efficiency; the pectinase used is fixed for easy separation and has high activity and high reuse rate.

[0006] In order to achieve the above object, in a first aspect, the present invention provides a fungal extracellular pectinase composition, which is composed of the following raw materials in parts by weight: 50-100 parts of pectinase composite particles, 0-10 parts of cellulase, 5-16 parts of acid amylase, and 0.5-1.5 parts of surfactant;

[0007] The pectinase composite particles are prepared by loading liposomes encapsulating fungal extracellular pectinase in chitosan gel.

[0008] Preferably, the preparation method of the pectinase composite particles is:

[0009] 1) Dissolving fungal extracellular pectinase, soybean lecithin and cholesterol in a solvent and subjecting the mixture to rotary evaporation to obtain liposomes encapsulating the fungal extracellular pectinase;

[0010] 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a buffer solution and subjecting the solution to electric field pulses to obtain porous activated liposomes;

[0011] 3) The chitosan acetic acid solution, the porous activated liposomes and the genipin crosslinking agent are mixed and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain pectinase composite particles.

[0012] Preferably, in step 1), the mass ratio of fungal extracellular pectinase, soybean lecithin and cholesterol is 0.1-1:5-20:0.1-5.

[0013] Preferably, the fungal extracellular pectinase comprises one or more of protopectinase, pectinesterase, pectin hydrolase or pectin lyase.

[0014] Preferably, the temperature of the rotary evaporation is 45-55°C.

[0015] Preferably, the buffer in step 2) is citric acid-sodium citrate buffer or phosphate buffer.

[0016] Preferably, the parameters of the electric field pulse in step 2) are: electric field intensity of 2 kV / cm to 6 kV / cm; and treatment time of 5 min to 10 min.

[0017] Preferably, in step 3), the volume mass ratio of chitosan acetic acid solution, porous activated liposomes, and genipin cross-linking agent is 100 mL: 1-10 g: 4-6 g.

[0018] Preferably, the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution at a volume-to-mass ratio of 1 g:5-8 mL.

[0019] Preferably, the surfactant is Tween-80 or Triton X-100.

[0020] In a second aspect, the present invention provides the use of the above-mentioned fungal extracellular pectinase composition in the preparation of fruit juice and vegetable juice.

[0021] The beneficial effects of the present invention include at least:

[0022] 1) The fungal extracellular pectinase composition obtained by the present invention has a synergistic effect of multiple enzymes and has a fast decomposition speed of pectin.

[0023] 2) The pectinase composite particles prepared by the present invention have high activity, high reusability, high environmental tolerance, and high activity at higher temperature and low pH.

[0024] 3) The fungal extracellular pectinase composition of the present invention has a simple preparation method, is naturally degradable, non-toxic, and is suitable for industrial production. DETAILED DESCRIPTION

[0025] The present invention provides a fungal extracellular pectinase composition, which is composed of the following raw materials in parts by weight: 50-100 parts of pectinase composite particles, 0-10 parts of cellulase, 5-16 parts of acid amylase, and 0.5-1.5 parts of surfactant;

[0026] The pectinase composite particles are prepared by loading liposomes encapsulating fungal extracellular pectinase in chitosan gel.

[0027] Preferably, the weight proportion of the raw materials is: 80 parts of pectinase composite particles, 3 parts of cellulase, 8 parts of acid amylase, and 1 part of surfactant. By adopting the above technical scheme, pectinase, cellulase, and acid amylase are used to cooperate with each other to accelerate the decomposition of pectin; cellulase can destroy the rigid structure of the cell wall and expose more pectin, so that pectinase can act on pectin more effectively, thereby accelerating the degradation process of pectin; adding acid amylase can specifically hydrolyze starch and glycogen. These polysaccharides will combine with pectin during the pectin extraction process to form a complex structure, which hinders the release and purification of pectin. By adding acid amylase, these polysaccharides can be decomposed, thereby releasing more pectin and improving the purity of the final product.

[0028] Preferably, the preparation method of the pectinase composite particles is:

[0029] 1) Dissolving fungal extracellular pectinase, soybean lecithin and cholesterol in a solvent and subjecting the mixture to rotary evaporation to obtain liposomes encapsulating the fungal extracellular pectinase;

[0030] 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a buffer solution and subjecting the solution to electric field pulses to obtain porous activated liposomes;

[0031] 3) The chitosan acetic acid solution, the porous activated liposomes and the genipin crosslinking agent are mixed and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain pectinase composite particles.

[0032] Preferably, in step 1), the mass ratio of fungal extracellular pectinase, soybean lecithin, and cholesterol is 0.1-1: 5-20: 0.1-5. By adopting the above technical solution, fungal extracellular pectinase can be encapsulated in liposomes, and the liposomes can form a relatively independent microenvironment, protecting the internal pectinase from the direct influence of adverse conditions such as extreme temperature, pH value and organic solvents, reducing the loss of enzyme activity, and allowing the enzyme to maintain activity under a wider range of processing conditions.

[0033] Preferably, the fungal extracellular pectinase includes one or more of protopectinase, pectinesterase, pectin hydrolase or pectin lyase; further preferably, a combination of protopectinase, pectinesterase, pectin hydrolase and pectin lyase in a mass ratio of 2:2:8:6. By adopting the above technical scheme, protopectinase can decompose the water-insoluble protopectin in the plant cell wall to release an enzyme of highly polymerized water-soluble pectin substances, which is conducive to separating water-soluble pectin from other substances by filtration to obtain water-soluble pectin; pectinesterase uses pectin with a methyl esterification degree of 65%-75% as a substrate to break the methyl ester bond of the methyl esterified pectin molecule; pectin hydrolase breaks the α-1,4 glycosidic bond of the main chain of the pectin polymer by hydrolysis; the role of pectin lyase is to break the α-1,4 glycosidic bond of the pectin polymer by trans-elimination reaction; the above pectinase compound can cope with a variety of pectins and can be widely used.

[0034] Preferably, the temperature of the rotary evaporation is 45-55°C.

[0035] Preferably, the buffer in step 2) is citric acid-sodium citrate buffer or phosphate buffer, more preferably citric acid-sodium citrate buffer.

[0036] Preferably, the parameters of the electric field pulse in step 2) are: electric field strength of 2 kV / cm~6 kV / cm; treatment time of 5 min~10 min. By adopting the above technical scheme, the liposome membrane can be perforated to increase permeability, which can not only protect pectinase but also increase the contact between pectinase and reaction substrate, thereby increasing the utilization rate of pectinase; and the present application found that the enzymatic hydrolysis of pectinase is enhanced under the treatment of the electric field pulse, and the pulsed electric field changes the conformation of pectinase, making its active center easier to bind to the substrate, exposing more active sites, and thus improving the catalytic efficiency of the enzyme.

[0037] Preferably, in step 3), the volume mass ratio of chitosan acetic acid solution, porous activated liposomes, and genipin crosslinker is 100mL: 1-10g: 4-6g. By adopting the above technical scheme, the present invention fixes the porous activated liposomes in the chitosan gel for easy separation and reuse, and chitosan is non-toxic, degradable, green and pollution-free, and can be used in the food industry; the role of genipin is as a crosslinker, which can react with the amino acid residues in the chitosan molecules, connect the chitosan molecules by forming covalent bonds, thereby constructing a three-dimensional network structure, and has safety and biocompatibility, can be safely used in the food field, and also has a certain substance transfer effect, increasing the reaction efficiency.

[0038] Preferably, the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution at a volume-to-mass ratio of 1 g:5-8 mL.

[0039] Preferably, the surfactant is Tween-80 or Triton X-100. By adopting the above technical solution, the present invention uses the surfactant to make the pectinase in the liposome contact with the reaction substrate better and increase the reaction efficiency.

[0040] In the second aspect, the present invention provides the use of the fungal extracellular pectinase composition in the preparation of fruit juice and vegetable juice.

[0041] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0042] It should be understood that the terms described in the present invention are only for describing special embodiments and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. Each smaller range between the intermediate value in any stated value or stated range and any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0043] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0044] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present application description and examples are exemplary only.

[0045] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0046] The "room temperature" and "normal temperature" mentioned in the present invention are all calculated as 25±2°C unless otherwise specified.

[0047] The raw materials used in the following examples of the present invention are all commercially available.

[0048] Example 1

[0049] Preparation of pectinase complex granules:

[0050] 1) Weigh the raw materials according to the mass ratio of fungal extracellular pectinase, soybean lecithin and cholesterol of 0.8:12:1, then dissolve them in chloroform and rotary evaporate them at 50°C to obtain liposomes encapsulating fungal extracellular pectinase; wherein the fungal extracellular pectinase is a combination of protopectinase, pectinesterase, pectin hydrolase and pectin lyase in a mass ratio of 2:2:8:6 (average enzyme activity 15000U / g).

[0051] 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a citric acid-sodium citrate buffer and subjecting the liposomes to an electric field pulse at an electric field strength of 4 kV / cm for 7 min to obtain porous activated liposomes;

[0052] 3) The chitosan acetic acid solution, the porous activated liposome, and the genipin crosslinker are mixed in a volume mass ratio of 100 mL: 5 g: 5 g and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain 50 μm pectinase composite particles; wherein the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution in a volume mass ratio of 1 g: 7 mL.

[0053] Preparation of fungal exocellular pectinase composition:

[0054] 80 parts of pectinase composite particles, 3 parts of cellulase, 8 parts of acid amylase and 1 part of Tween-80 were weighed according to the mass ratio, stirred and mixed evenly to obtain a fungal extracellular pectinase composition.

[0055] Example 2

[0056] Preparation of pectinase complex granules:

[0057] 1) Weigh the raw materials according to the mass ratio of fungal extracellular pectinase, soybean lecithin and cholesterol of 0.1:5:0.1, then dissolve them in chloroform and rotary evaporate them at 45°C to obtain liposomes encapsulating fungal extracellular pectinase; wherein the fungal extracellular pectinase is a combination of protopectinase, pectinesterase, pectin hydrolase and pectin lyase in a mass ratio of 2:2:8:6 (average enzyme activity 15000 U / g).

[0058] 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a citric acid-sodium citrate buffer and subjecting the liposomes to an electric field pulse at an electric field strength of 2 kV / cm for 5 min to obtain porous activated liposomes;

[0059] 3) The chitosan acetic acid solution, the porous activated liposome, and the genipin crosslinker are mixed in a volume mass ratio of 100 mL: 1 g: 4 g and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain 100 μm pectinase composite particles; wherein the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution in a volume mass ratio of 1 g: 5 mL.

[0060] Preparation of fungal exocellular pectinase composition:

[0061] Weigh 50 parts of pectinase composite particles, 1 part of cellulase, 10 parts of acid amylase, and 0.5 parts of Tween-80 according to the mass ratio, stir and mix evenly to obtain a fungal extracellular pectinase composition.

[0062] Example 3

[0063] Preparation of pectinase complex granules:

[0064] 1) Weigh the raw materials according to the mass ratio of fungal extracellular pectinase, soybean lecithin and cholesterol of 1:20:5, then dissolve them in chloroform and rotary evaporate them at 55°C to obtain liposomes encapsulating fungal extracellular pectinase; wherein the fungal extracellular pectinase is a combination of protopectinase, pectinesterase, pectin hydrolase and pectin lyase in a mass ratio of 2:2:8:6 (average enzyme activity 15000U / g).

[0065] 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a phosphate buffer solution and subjecting the solution to an electric field pulse at an electric field strength of 6 kV / cm for 10 min to obtain porous activated liposomes;

[0066] 3) The chitosan acetic acid solution, the porous activated liposome, and the genipin crosslinker are mixed in a volume mass ratio of 100 mL: 10 g: 6 g and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain 10 μm pectinase composite particles; wherein the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution in a volume mass ratio of 1 g: 8 mL.

[0067] Preparation of fungal exocellular pectinase composition:

[0068] 100 parts of pectinase composite particles, 10 parts of cellulase, 16 parts of acid amylase and 1.5 parts of Triton X-100 were weighed according to the mass ratio, stirred and mixed evenly to obtain a fungal extracellular pectinase composition.

[0069] Example 4

[0070] The preparation method of this embodiment is the same as that of embodiment 1, except that the fungal extracellular pectinase is a combination of protopectinase and pectin hydrolase in a mass ratio of 2:8 (average enzyme activity 15000 U / g).

[0071] Example 5

[0072] The preparation method of this embodiment is the same as that of embodiment 1, except that the fungal extracellular pectinase is pectin hydrolase (average enzyme activity 15000 U / g).

[0073] Example 6

[0074] The preparation method of this embodiment is the same as that of embodiment 1, except that cellulase is not contained therein.

[0075] Comparative Example 1

[0076] Preparation of pectinase complex granules:

[0077] The chitosan acetic acid solution, pectinase combination, and genipin crosslinker are mixed in a volume mass ratio of 100 mL: 0.8 g: 5 g and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain 10 μm pectinase composite particles; wherein the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution in a volume mass ratio of 1 g: 8 mL. The pectinase combination is a combination of protopectinase, pectinesterase, pectin hydrolase, and pectin lyase in a mass ratio of 2: 2: 8: 6 (average enzyme activity 15000 U / g).

[0078] Preparation of fungal exocellular pectinase composition:

[0079] 100 parts of pectinase composite particles, 10 parts of cellulase, 16 parts of acid amylase and 1.5 parts of surfactant were weighed according to the mass ratio, stirred and mixed evenly to obtain a fungal extracellular pectinase composition.

[0080] Comparative Example 2

[0081] Preparation of fungal exocellular pectinase composition:

[0082] 80 parts of pectinase combination, 10 parts of cellulase, 16 parts of acid amylase and 1.5 parts of surfactant were weighed according to the mass ratio, stirred and mixed evenly to obtain a fungal extracellular pectinase composition.

[0083] Comparative Example 3

[0084] The preparation method of this embodiment is the same as that of embodiment 1, except that the electric field pulse treatment is not performed when the pectinase composite particles are prepared.

[0085] Comparative Example 4

[0086] Preparation of pectinase complex granules:

[0087] 1) Weigh the raw materials according to the mass ratio of fungal extracellular pectinase, soybean lecithin and cholesterol of 0.8:12:1, then dissolve them in chloroform and rotary evaporate them at 50°C to obtain liposomes encapsulating fungal extracellular pectinase; wherein the fungal extracellular pectinase is a combination of protopectinase, pectinesterase, pectin hydrolase and pectin lyase in a mass ratio of 2:2:8:6 (average enzyme activity 15000U / g).

[0088] 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a citric acid-sodium citrate buffer and ultrasonicating for 5 min at an ultrasonic power of 360 W with a frequency of 5 s and a rest of 3 s to obtain porous liposomes;

[0089] 3) The chitosan acetic acid solution, porous liposomes, and genipin crosslinker are mixed in a volume mass ratio of 100 mL: 5 g: 5 g and the pH is adjusted to neutral to obtain a gel material, and then the gel material is freeze-dried and crushed to obtain 50 μm pectinase composite particles; wherein the chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution in a volume mass ratio of 1 g: 7 mL.

[0090] Preparation of fungal exocellular pectinase composition:

[0091] 80 parts of pectinase composite particles, 3 parts of cellulase, 8 parts of acid amylase and 1 part of Tween-80 were weighed according to the mass ratio, stirred and mixed evenly to obtain a fungal extracellular pectinase composition.

[0092] Comparative Example 5

[0093] The preparation method of this embodiment is the same as that of embodiment 1, except that the cross-linking agent in step 3) is terephthalaldehyde, the concentration of terephthalaldehyde is 0.3%, and the amount used is 5 mL.

[0094] Experimental Example 1

[0095] Activity detection of pectinase composite particles

[0096] The pectinase composite particles of Examples 1-3 and Comparative Examples 1 and 3 were tested for enzyme activity recovery rate using the sodium hypoiodite method, and the enzyme activity after treatment at different temperatures and pH values ​​was tested. The test results are shown in Table 1.

[0097] Table 1

[0098]

[0099] It can be seen from Table 1 that the pectinase composite particles prepared in the present invention have little effect on enzyme activity during the preparation process and have resistance to the external environment; combined with Example 1, Comparative Example 3 and Comparative Example 4, it is illustrated that the electric field pulse has a positive effect on the activity of pectinase liposomes. Comparative Example 3 shows that the construction of liposomes has a protective effect on pectinase but has a certain effect on the utilization rate of the enzyme.

[0100] Experimental Example 2

[0101] Test on enzymatic hydrolysis of pear juice by fungal extracellular pectinase combination

[0102] The pear pulp was crushed and juiced, and 5 mL of juice was evenly taken into test tubes. 1 g of the fungal extracellular pectinase composition of each embodiment or comparative example was added to each test tube. Then, the pear juice was enzymolyzed at 50° C. and the transmittance of the pear juice was measured at 660 nm after 1 h, 2 h, and 3 h, as shown in Table 2.

[0103] Table 2

[0104]

[0105] It can be seen from Table 2 that the fungal extracellular pectinase composition obtained in the present invention has a strong ability to decompose pectin and has a rapid decomposition effect.

[0106] Experimental Example 3

[0107] Pectinase composite particles circulation activity detection

[0108] The pectinase composite particles of Examples 1-3 and Comparative Examples 1 and 3 were used to hydrolyze pears for 3 h using the method of Experimental Example 2. This was repeated 50 times, and the enzyme activity was tested. See Table 3.

[0109] Table 3

[0110] Group Enzyme activity recovery rate%% Example 1 82.3 Example 2 78.6 Example 3 80.7 Comparative Example 1 48.3 Comparative Example 3 53.8

[0111] It can be seen from Table 3 that the pectinase composite particles obtained in the present invention still have high activity after 50 cycles of use and have a high reuse rate.

[0112] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. A fungal extracellular pectinase composition, characterized in that: The method is composed of the following raw materials in parts by weight: 50-100 parts of pectinase composite particles, 0-10 parts of cellulase, 5-16 parts of acid amylase, and 0.5-1.5 parts of surfactant; The pectinase composite particles are prepared by loading liposomes encapsulating fungal extracellular pectinase in chitosan gel; The preparation method of the pectinase composite particles is as follows: Step 1) dissolving fungal extracellular pectinase, soybean lecithin and cholesterol in a solvent and subjecting the solvent to rotary evaporation to obtain liposomes encapsulating the fungal extracellular pectinase; Step 2) dissolving the liposomes encapsulating the fungal extracellular pectinase in a buffer solution and subjecting the solution to electric field pulses to obtain porous activated liposomes; Step 3) mixing the chitosan acetic acid solution, the porous activated liposomes, and the genipin crosslinker and adjusting the pH to neutral to obtain a gel material, and then freeze-drying and crushing the gel material to obtain pectinase composite particles; The parameters of the electric field pulse in step 2) are: electric field intensity of 2 kV / cm to 6 kV / cm; and treatment time of 5 min to 10 min.

2. The fungal extracellular pectinase composition according to claim 1, characterized in that In the step 1), the mass ratio of fungal extracellular pectinase, soybean lecithin and cholesterol is 0.1-1:5-20:0.1-5.

3. The fungal extracellular pectinase composition according to claim 2, characterized in that The fungal extracellular pectinase comprises one or more of protopectinase, pectinesterase, pectin hydrolase or pectin lyase.

4. The fungal extracellular pectinase composition according to claim 1, characterized in that The buffer in step 2) is citric acid-sodium citrate buffer or phosphate buffer.

5. The fungal extracellular pectinase composition according to claim 1, characterized in that In the step 3), the volume mass ratio of the chitosan acetic acid solution, the porous activated liposomes, and the genipin cross-linking agent is 100 mL: 1-10 g: 4-6 g.

6. The fungal extracellular pectinase composition according to claim 5, characterized in that The chitosan acetic acid solution is prepared by mixing chitosan powder and 2% acetic acid solution at a volume mass ratio of 1g:5-8mL.

7. The fungal extracellular pectinase composition according to claim 1, characterized in that The surfactant is Tween-80 or Triton X-100.

8. Use of the fungal extracellular pectinase composition according to any one of claims 1 to 7 in the preparation of fruit juice or vegetable juice.

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